Total RNA Kit, peqGOLD

RNA isolation / purification Tissue - Human Stomach

Experiment
RNA isolation / purification Tissue - Human Stomach
Product
Total RNA Kit, peqGOLD from VWR
Manufacturer
VWR

Protocol tips

Upstream tips
- This kit purifies RNA from up to 100ug tissue
Protocol tips
- All steps must be carried out at room temperature (22 – 25 °C).
- Pre-heat RNase-free Water to 70 °C prior to elution.

- For DNA contamination Digest with Rnase free DNase and incubate at 37 °C for 5 min.
Downstream tips
- Ensure RNA Wash Buffer II Concentrate has been diluted with 4 volumes of 100 % ethanol as indicated on the bottle for ant salt carryover during elution

Publication protocol

Total RNA was extracted from HGT-1 cells and two human biopsy specimens by using the peqGold Total RNA Kit (Peqlab). Quantity and quality were checked spectrophotometrically. Reverse transcription was carried out with 2 µg RNA and the High Capacity cDNA Reverse Transcription Kit (Thermo Fisher Scientific). Real-time PCR was performed with an Applied Biosystems StepOneplus Real Time PCR system and Fast SYBR Green Master Mix (Thermo Fisher Scientific). Primers were designed using the National Center for Biotechnology Information (NCBI) primer designing tool (using Primer 3 and BLAST) . Cycling conditions were 20 s/95 °C (activation), 3 s/95 °C (denaturation), 30 s/60 °C (annealing), and 15 s/67 °C (elongation with fluorescence measurement)

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Reviews

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Discussion

Discussion

4 years ago

Author: Paul G. Macon United States

RNA isolation from tissue

How do I extract RNA from animal tissue without using liquid nitrogen? I tried the RNA extraction by using the TRIzol reagent and I homogenize the tissue using polytron homogenizer at room temperature for 30secs is this correct?

Discussion

5 years ago

Author: Aaron Stege Netherlands

Problem in phase separation after using serum/plasma kit

I used a serum/plasma kit for my serum samples. After the phase separation the samples should have 3 phases: a colourless aqueous phase, a white interphase and a red organic phase. However, in some of my samples there was no aqueous phase unless I wait for an extended period of time. How can I circumvent this problem?

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing RNA isolation / purification Tissue - Human Stomach using Total RNA Kit, peqGOLD from VWR.

Paper title
Caffeine induces gastric acid secretion via bitter taste signaling in gastric parietal cells
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Manufacturer protocol

Download the product protocol from VWR for Total RNA Kit, peqGOLD below.

Download PDF Download manufacturer protocol

Videos

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